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J R Vogt

Publications and source records attributed to J R Vogt.

8 recordsLinked to original sources

Biosynthesis of vitamin B12 in anaerobic bacteria. Experiments with Eubacterium limosum on the incorporation of D-[1-13C]erythrose and [13C]formate into the 5,6-dimethylbenzimidazole moiety.

Experiments on the incorporation of erythrose and formate into the 5,6-dimethylbenzimidazole moiety of vitamin B12 are described. In one experiment, a 1:1 mixture of D-[1-13C]erythrose and D-[1-13C]threose was added to a Eubacterium limosum fermentation. The vitamin B12 formed was methylated at N3 of its 5,6-dimethylbenzimidazole part and degraded to 1,5,6-trimethylbenzimidazole. The 13C-NMR spectrum of this compound exhibited a single prominent signal at 109.5 ppm due to 13C labeling in C7. This shows that C1 of erythrose or threose was originally incorporated exclusively into C4 of the 5,6-dimethylbenzimidazole moiety of vitamin B12. In another experiment, sodium [13C]formate was added to a culture of E. limosum. The vitamin B12 isolated was transformed into 1,5,6-trimethylbenzimidazole as before. The 13C-NMR spectrum also showed one prominent signal at 142.8 ppm, evoked by 13C at C2. These results demonstrate that erythrose is incorporated into the base part of vitamin B12 regiospecifically and that formate is the precursor of the C2.

Benzimidazoles↗

Biosynthesis of vitamin B-12 in anaerobic bacteria. Experiments with Eubacterium limosum and D-erythrose 14C-labeled in different positions.

In anaerobic microorganisms the origin of C atoms 2 and 4-7 of the 5,6-dimethylbenzimidazole moiety of vitamin B-12 is still unknown. In order to tackle this problem we added several 14C-labeled putative precursors to Eubacterium limosum fermentations. The degradation of the isolated vitamin B-12 revealed that only D-erythrose, 14C-labeled in different positions, was efficiently incorporated into the 5,6-dimethylbenzimidazole part. The 5,6-dimethylbenzimidazole obtained from an experiment with D-[U-14C]erythrose was further degraded. It was found that C-2 was unlabeled, whereas half of the label was located in C-5 plus C-6, and the other half in C-4 plus C-7. These results demonstrate that in E. limosum D-erythrose is a precursor of C-atoms 4, 5, 6 and 7 of the 5,6-dimethylbenzimidazole part of vitamin B-12.

Eubacterium↗

Biosynthesis of vitamin B-12 in anaerobic bacteria. Experiments with Eubacterium limosum on the origin of the amide groups of the corrin ring and of N-3 of the 5,6-dimethylbenzimidazole part.

The pathway of vitamin B-12 biosynthesis in anaerobic bacteria differs in several respects from the pathway found in aerobic or aerotolerant microorganisms. The aim of this investigation was to elucidate the formation of the 5,6-dimethylbenzimidazole part and the amide groups of vitamin B-12 in anaerobic bacteria. [15N]Ammonium chloride or L-[amido-15N]glutamine or a mixture of [15N]ammonium sulfate and [15N]glycine was added to fermentations with Eubacterium limosum. The vitamin B-12 isolated from these fermentations was methylated and degraded to cobinamide and 1,5,6-trimethylbenzimidazole. The amide groups of cobinamide were hydrolyzed and the amide nitrogen of the side chains a, b, c, d, e and g trapped as benzamide. The 15N incorporation was determined by mass spectroscopy. Thus in the experiment with [15N]ammonium chloride the benzamide and the 1,5,6-trimethylbenzimidazole contained 9.6% 15N, whereas in the experiment with L-[amido-15N]glutamine 37.5% of the molecules were 15N labeled. The 1H-NMR spectrum of 1,5,6-trimethylbenzimidazole revealed that the 15N from the ammonium salts and from glutamine was incorporated into N-3 of the 5,6-dimethylbenzimidazole moiety of vitamin B-12. With a mixture of [15N]ammonium sulfate and [15N]glycine both nitrogens of 5,6-dimethylbenzimidazole became 15N-labeled. These experiments demonstrate that in E. limosum the amide nitrogen of glutamine is not only the precursor of the six amide groups of the corrin ring, but also of N-3 of the 5,6-dimethylbenzimidazole moiety of vitamin B-12.

Amides↗

Feasibility studies in rats fed heavy metals as multiple nutrient markers.

The nitrates of five metals (cerium, terbium, ytterbium, lutetium, and iridium) were fed to rats to determine the feasibility of their use as nonabsorbed, multiple markers for recovery, passage, and indirect apparant digestibility studies. Fecal recovery of a single oral dose was completed within 72 hours. When the salts were mixed into the diet, 48-96 hours was required to establish a steady-state concentration of markers in feces. The diurnal variation of cerium in feces was found to be considerable when it was fed twice daily as a single dose prior to each feeding. When incorporated into the diet, negligible diurnal variation in fecal concentration was noted with lutetium, and small variation was seen with other metals. In nutrient apparent digestibility studies, good agreement was generally found between direct and indirect multiple marker methods. Experiments with a daily intake marker suggest that cerium was not satisfactory as a multiple marker in which neutron activation analysis was the method of determination.

Animal Nutritional Physiological Phenomena↗